Abstract Worldwide, Non-Small Cell Lung Cancer (NSCLC) is the most common and among the most lethal of human cancers with about 2 million new cases per year and a 5 year survival for metastatic disease of about 1%. Fortunately, a number of new treatments have improved the lives of some patients with NSCLC. During the past decade, the Moffitt Thoracic Oncology Department has pioneered new strategies using immunotherapy for NSCLC. In almost 500 patients treated with immunotherapy at Moffitt, the response rate (CR, PR, and SD) is 30 to 45% (2, 3). Most responses are followed by evolution of resistance and progression but some patients in each category have experienced durable responses maintained for > 1 year (2). Our underlying hypothesis is that the observed results from immunotherapy can be improved with sufficient understanding of the evolutionary (cellular and molecular) and ecological (tissue) dynamics that govern response and resistance of NSCLC to immunotherapy. We have previously demonstrated that administration of cancer therapy can be optimized through evolutionary mathematical models that frame the complex, often non-linear underlying dynamics. To develop such models in immunotherapy of NSCLC, we will analyze a Moffitt NSCLC immunotherapy patient cohort all of whom were treated within an investigational protocol in which tumor biopsies are performed prior to therapy and after 6 weeks of immunotherapy. They also underwent repeated imaging with radiomic analyses and blood studies during the course of therapy, and many had biopsies at the time of progression. Retrospective analysis of this cohort will investigate the evolutionary (molecular, cellular) data as well as ecological (histological and radiological) dynamics that govern response and resistance to immunotherapy. These investigations will be supplemented from additional ex vivo studies in which tumor and immune cells obtained from resected primary tumors are dispersed in culture allowing the immediate response to immunotherapy agents to be assessed. We will also perform in vitro studies that dissect the wide range of cellular and tissue ecological engineering strategies available to NSCLC cells as well as the timescales of immunotherapy adaptation. Finally, we will test the predictive power of our developed mathematical models to use pre-therapy data to predict outcomes from monotherapy with PD-L1 checkpoint inhibitors. We will then extend these models by integrating additional immunosuppressive mechanisms and test these models in a second clinical cohort treated with combinations of PD-LI and CTLA-4 checkpoint inhibitors.